For the first time in the world! Sperm produced in vitro by sperm cells survive for 800 days Provide new ideas for the protection of endangered fish and the optimization and breeding of economic fish

Recently, Professor Li Mingyou and Associate Professor Gui Lang’s team from the School of Fisheries and Life Sciences of Shanghai Ocean University have made progress in the research on in vitro induction of spermatogenesis in fish spermatogon stem cell lines (SSCs), and long-term stable culture of sarcoma sperm stem cell lines can be induced by fish gonadal cells to generate sperm. The study has been published in Biology.


The “magical” transformation of stomato seminal stem cells courtesy of the interviewee

This is the world’s first long-term cultivation of sperm stem cell lines for economic fish, which can not only produce sperm in vitro, but also survive for up to 800 days, which provides a basis for further research and understanding of sperm stem cells, and also provides new ideas for protecting endangered fish species and optimizing the breeding of fish of important economic value.

The salmon is a small economic fish, from a small and round stuccos sperm stem cells, to the sperm that can develop free swing of the flagella in the experimental dish, the reproductive stem cells of long-term cultivated economic fish have completed the magical journey of developing sperm in the culture medium, which is the first time in the world.

Spermatogenesis and meiosis produce sperm, passing on genetic information to the next generation, thus ensuring the continuation of the species. Numerous studies have shown that the remodeling spermatogenesis microenvironment in vitro can induce spermatogonocyte line differentiation to produce sperm. In fish research, obtaining spermatocyte lines in vitro and being able to culture them for a long time is a key technical challenge.

Research institutions around the world are exploring spermatocyte lines that can be reliably cultured over a long period of time, but usually only fish sperm stem cell lines can be obtained for up to a month. On the basis of the research results of their predecessors, Li Mingyou and Guilang’s team applied the model fish stem cell culture technology to the salmon and knife fish, and established the first long-term culture of farmed fish in vitro in the world – the spermogen stem cell line of the salmon.

Sarcoma spermatonchynocytes differentiate into three germ layers of different cells in zebrafish.png

Differentiation of spermatogenic stem cells in zebrafish Courtesy of the interviewee

“Our sarcoma spermatose stem cell line has survived in vitro for more than two years, which should be the longest surviving time for farmed fish in the world, which is the ‘Guinness’ record on the spermatogon stem cell line.” Li Mingyou said, “And its growth is very stable, with a typical spermatocyte gene expression pattern.” As a result, a long-term cultured and sperm-producing sperm stem cell line was established, as well as a Yangtze River knife scorpion glandular cell line that can induce sperm production in other fish sperm stem cell lines.

In the laboratory, the sarcoma spermatogenesis stem cell line is co-cultured with the stomatose sperm nest cells, and the cells can enter and complete meiosis to produce haploid products. By day 14, sperm cells expressing red fluorescent protein were observed in the medium, indicating that the mammoth seminal stem cell line was gradually induced into immature sperm cells. After 16 days of culture, the spermatozoogen stem cell lines of the horsefish were observed to differentiate into more mature sperm with a slight wobbling behavior in the tail. This suggests that under suitable induction conditions, the stingray spermatogenesis stem cell line can enter the spermatogenesis phase, producing “magical” sperm cells.

The experimenters transferred the spermatogen stem cell line of the horsefish into the zebrafish embryo, and they could be observed to differentiate into different cells derived from the three germ layers. In addition, it was also found that the spermatophile stem cell line of the marquee fish can colonize the zebrafish gonads and produce sperm.

At the same time, another study proved that somatic cells play an important role in the process of spermatogenesis, and can induce in vitro differentiation of sperm stem cell lines by simulating the in vivo microenvironment by using somatic cells and spermatogonial stem cell line co-culture conditions, and undergo meiosis to produce sperm cells.

This study broke through the problem of long-term cultivation of farmed fish sperm stem cell lines, and through multi-angle induction and transplantation studies inside and outside the body, sperm was formed in vitro of sperm. This provides a basis for further research and understanding of spermatocyte lines, as well as new ideas for the protection of endangered fish species and the optimization of the breeding of fish of economic importance.

In the next step, the team will also combine stem cell culture and induction technology, gene editing technology and cell transplantation technology to establish reproductive stem cell systems for rare or endangered fish with difficult reproduction, long reproductive cycle, and carry out new technologies for the creation of excellent germplasm of cultured fish mediated by reproductive stem cell gene editing. (Source: China Science Daily, Zhang Shuanghu, Huang Xin)

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